International partners remain deeply troubled after Trump's campaign rhetoric questioned long-established alliances, and they worry about a growing scandal over possible ties between some of Trump's staff and Moscow.
"The transatlantic bond remains our strongest bulwark against instability and violence," Mattis told the Munich Security Conference.
"I am confident that we will strengthen our partnerships, confronting those who choose to attack innocent people or our democratic processes and freedoms."
Trump has said he is open to closer cooperation with Moscow on Syria, particularly in the fight against the Islamic State jihadist group.
But Mattis told a NATO summit yesterday that he is not "in a position right now to collaborate on a military level" with Moscow and said Russia must first "prove itself" and follow international law before envisioning any closer military ties.
"European leaders and European governments are leaders are extremely impatient to find out what will really drive US foreign policy in this new period," he said, underscoring that many in Europe are clamouring to find out about the future of US-Russian relations.
The White House is still reeling from the forced resignation Monday of Trump's national security advisor, Michael Flynn, after he held pre-inauguration phone calls with the Russian ambassador about US sanctions policy.
Trump, who has repeatedly praised Russian President Vladimir Putin, yesterday denied he or his staff had any pre-election contacts with Moscow.
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